Pre-zygotic breeding barriers between the wild diploid potato species Solanum maglia and S. kurtzianum from Argentina

Solanum maglia Schldl. is a wild tuber-bearing species from the Quebrada de Alvarado (Mendoza, Argentina) and the Valparaíso area (Chile). Only diploid cytotypes are known from Argentina, while diploids and triploids are found in Chile. Five diploid accessions from Argentina did not produce seeds upon intercrossing. Thus, the basis of this behaviour and the crossability relations of this species with S. kurtzianum, Bitt. et Wittm., another diploid species that grows in its vicinity were investigated. After checking pollen viability, intraspecific and interspecific pollinations were carried out following an incomplete diallel mating design. Five emasculated flowers were pollinated per each one of 135 different combinations of genotypes (crosses); three flowers were fixed in FAA and analyzed under fluorescent microscopy, whereas the remaining two were left on the plant for fruit and seed set. Pollen tube growth was arrested in the first third of the style and no seeds were obtained in intraspecific S. maglia crosses. In interspecific crosses and according to their direction, pollen tubes either grew normally to the base of the style or were arrested in the last third, producing no fruits or fruits with a variable number of seeds in each. It is concluded that the five S. maglia accesions are, possibly, one clone and, thus, the gametophytic auto-incompatibility system prevents seed formation upon intercrossing. On the other hand, the incompatibility reaction observed in the interspecific crosses might be controlled by one or more genes independent of the S-locus.     

Crossability relationships between the common potato, Solanum tuberosum spp. tuberosum, and its wild diploid relatives S. kurtzianum and S. ruiz-lealii

The common potato, Solanum tuberosum spp. tuberosum (tbr), is a tetraploid species with a narrow genetic base, but with a large number of related species that harbor great genetic diversity for numerous characters of agronomic interest. S. kurtzianum (ktz) and S. ruiz-lealii (rzl) are wild diploid relatives with potential resistance to adverse biotic and abiotic factors. To evaluate if this wild germplasm can be incorporated by conventional crosses into the tbr gene pool, pollen-pistil compatibility relations and seed set in interspecific crosses with tbr were investigated. In 4x tbr × 2x ktz crosses and the reciprocals, 89% and 52.2% of genotypic combinations (respectively) were compatible at the pollen-pistil level. Seeds were obtained from some genotypic combinations in both directions of the cross, suggesting that functional 2n gametes might be produced by particular ktz genotypes. In 2x rzl × 4x tbr crosses and their reciprocals, 35% and 11.7% (respectively) of the genotypic combinations were compatible at the pollen-pistil level, but no seeds were obtained. These results indicate that the reproductive isolation between 4x tbr and 2x ktz is incomplete, and that gene exchange between them is feasible. Moreover, further studies will have to be carried out with rzl to ascertain its crossability with 4x tbr.     

Isoenzyme differences between the wild diploid and tetraploid wheats

Variability of acid phosphatase (ACP), esterase (EST), and superoxide dismutase (SOD) isoenzymes among 660 accessions of wild diploid wheats Triticum boeoticum (475) and T. urartu (185) and among 35 accessions of wild tetraploid wheats T. araraticum (21) and T. dicoccoides (14) was studied with the use of polyacrylamide gel electrophoresis. The wild diploids are differentiated by the electrophoretic variants of ACP-B, ACP-C, EST-A, EST-B, EST-C, EST-D, and SOD-B. Triticum urartu matches the A genome of wild tetraploid wheats T. araraticum and T. dicoccoides by allozymes of ACP-B, ACP-C and SOD-B, but differs distinctly by the monomorphic allozyme EST-A66. Triticum boeoticum fits the A genome of polyploid wheats by allozymes of EST-A and EST-B, but differs in allozymes of ACP-B and SOD-B. Thus, none of the two contemporary diploid wheats corresponds exactly to the A genome of wild tetraploid wheats by all diagnostic isoenzymes. Triticum boeoticum and T. arararicum share EST-C46 and EST-D71, whereas T. urartu and T. dicoccoides are fixed for silent EST-C0 and share EST-D73, suggesting independent origin of the two tetraploid wheats with a contribution from ancestral forms of both diploids.     

Cytological mechanisms of 2n pollen formation in the wild potato Solanum okadae and pollen-pistil relations with the cultivated potato, Solanum tuberosum

Solanum okadae Hawkes et Hjert is a wild diploid potato species endemic to Argentina and Bolivia, of potential breeding value. However, no genetic studies have been carried out with this species and its crossability relations with the common tetraploid potato, S. tuberosum L. ssp. tuberosum, are unknown. Furthermore, accessions from both countries differed in their morphological phenotypes. To ascertain the feasibility of incorporating this wild germplasm into cultivated potato, 2n pollen screening was carried out in 10 accessions and families derived from crosses between accessions; also, pollen-pistil compatibility relations were studied in reciprocal interspecific crosses. Plants of four of the 10 accessions produced 2n pollen (0.1 to 5.0%) and 4n pollen (0.0 to 3.0%). Parallel and tripolar spindles at Anaphase II were the cytological mechanisms involved in 2n pollen formation; lack of chromosome migration in both meiotic divisions originated the 4n pollen. Both full compatibility and incompatibility at various sites along the pistil were observed in the S. tuberosum × S. okadae combinations; most reciprocal combinations were incompatible. Compatible genotypes produced 2n pollen. However, only a few seeds were obtained and chromosome counts could not be carried out in the hybrid progeny because seedlings died at an early stage. The pollen-pistil barriers are incomplete and can be circumvented by the appropiate choice of parents. The identification of the post-zygotic barriers will be the focus of further studies.     

Inheritance of morphological characters and glycoalkaloids in potatoes of somatic hybrids between dihaploid Solanum acauleand tetraploid Solanum tuberosum.

Steroidal glycoalkaloids occur in potatoes and are reported to impart resistance to phytopathogens including bacteria, fungi, and insects. Because glycoalkaloids can be passed to progenies during breeding programs designed to develop improved potatoes, it is of importance to determine the quality of desired characteristics and the composition of glycoalkaloids of new somatic hybrids. The objective of this study was to determine the appearance, size, and shape (morphological characters) as well as the glycoalkaloid content of potato tubers of somatic hybrids between tetraploid Solanum tuberosum cv. Dejima (2n = 4x = 48 chromosomes) and the dihaploid clone ATDH-1 (2n = 2x = 24 chromosomes) induced by anther culture from Solanum acuale-T (acl-T, 2n = 4x = 48 chromosomes). Tuber size and shape in somatic hybrids were in accord with those of cv. Dejima, whereas the tuber skin color resembled that of ATDH-1. Thin-layer chromatography, high-performance liquid chromatography, and gas-liquid chromatography/mass spectrometry studies showed that the two steroidal glycoalkaloids (alpha-chaconine and alpha-solanine) were present in the tubers of S. tuberosum, whereas acl-T and ATDH-1 tubers were found to contain alpha-tomatine and demissine. The concentrations of total glycoalkaloids in both acl-T and ATDH-1 was >100 mg/100 g of fresh weight tuber cortex, much higher than in S. tuberosum. All somatic hybrids, except one clone, contained four glycoalkaloids (alpha-chaconine, alpha-solanine, alpha-tomatine, and demissine) derived from the fusion parents. The lack of alpha-tomatine in the remaining clone may be due to somaclonal variation. The results show that character expression is influenced by ploidy level and that total glycoalkaloid levels in most somatic hybrids were intermediate between those of the fusion parents. The possible significance of these findings for plant breeding and food safety is discussed.     

Comparison of the calystegine composition and content of potato sprouts and tubers from Solanum tuberosum Group Phureja and Solanum tuberosum Group Tuberosum

The calystegines detected in tubers from 17 Phureja ( S. tuberosum Group Phureja) lines and five Tuberosum ( S. tuberosum Group Tuberosum) cultivars were identified as the A 3 and B 2 structural types. Their concentration in whole tubers was of a similar order of magnitude in both species, as was the variability in the ratio of B 2 to A 3. On average, calystegine concentrations in the peel were about 13 times that found in the flesh for the five Tuberosum cultivars, and 4 times higher for four Phureja lines. Removal of the peel reduced the calystegine content by an average of over 50% in Tuberosum but by only 30% in Phureja, despite the latter having the greater proportion of peel. The calystegine content of sprouts was also determined for five Tuberosum cultivars and four Phureja lines and was found to include small amounts of four additional types, B 3, B 4, N 1, and X 2, in addition to the more abundant A 3 and B 2. Concentrations in the sprouts of Tuberosum were on average 100 times higher than that in the tuber flesh and 8 times higher than in the peel, whereas for Phureja, the equivalent values were 30 and 7 times higher, respectively. No correlation was found between sprout concentration and either flesh or peel calystegine concentration.     

LC-MS analysis of solanidane glycoalkaloid diversity among tubers of four wild potato species and three cultivars (Solanum tuberosum)

Secondary metabolites in potato tubers include both phytonutrients and plant defense compounds. The extent these small molecules vary among different potato genotypes is not well characterized. LC-MS analysis of tuber extracts from seven potato genotypes showed that one large source of small molecule variation is the glycoalkaloids. Glycoalkaloids are involved in the resistance of potatoes to pathogens and pests, but they also have implications for human health and nutrition. This study focused on glycoalkaloids with solanidane or solanidane-like aglycones, of which over 50 were tentatively identified, many of which appeared to be novel glycoalkaloids. Results suggested the variety of glycoalkaloids in potatoes is considerably greater than previously thought. Dissecting the role of these many glycoalkaloids in human health or pest and pathogen resistance will be a formidable undertaking.     

Radical scavenging capacity and antimutagenic properties of purified proteins from Solanum betaceum fruits and Solanum tuberosum tubers

In this study, antioxidant activities in free-radical-mediated oxidative systems and the genotoxic/antigenotoxic effects of two proteins with molecular mass around 17 kDa, purified from Solanum betaceum fruits (cyphomine) and Solanum tuberosum tubers (solamarine), were investigated. Both proteins inhibited uric acid formation with IC(50) values between 55 and 60 μg/mL, and both proteins were able to reduce oxidative damage by scavenging hydroxyl radicals and superoxide anion in a dose-dependent manner. Furthermore, the DPPH? reduction assay showed SC(50) values of 55-73 μg/mL. Cyphomine and solamarine were able to retain their antioxidant activity after heat treatment at 80 °C for 15 min. Allium cepa and Salmonella /microsome assays showed no genotoxic and mutagenic effects. Solamarine showed an antimutagenic effect against a direct mutagen (4-nitro-o-phenylenediamine). Consequently, the present study showed that the investigated proteins are promising ingredients for the development of functional foods with a beneficial impact on human health and an important source for the production of bioactive peptides.     

Comparison of fatty acid and polar lipid contents of tubers from two potato species, Solanum tuberosum and Solanum phureja

Total and individual fatty acid contents were determined in raw tubers of four genotypes from each of the two species Solanum phureja and S. tuberosum. The four S. phureja genotypes contained statistically significantly higher concentrations of total fatty acids, with their average value being 37% greater than that for the mean of the four S. tuberosum cultivars. In both species, a total of 17 fatty acids were detected in quantifiable amounts, and in all genotypes the predominant fatty acid was linoleic followed by alpha-linolenic and palmitic acids. Unusually, 15-methyl hexadecanoate was present as a minor acid in both species. Although a number of statistically significant differences in the fatty acid percentage compositions were found between and within the two species, these were generally small. Averaged over all species and genotypes, tuber storage resulted in an initial small but statistically significant increase in total fatty acid content, but prolonged storage resulted in a fall to the initial values detected close to harvest. The same trend was evident for S. phureja alone (for mean values of all genotypes), but for S. tuberosum the total fatty acid content remained constant over the whole storage period. For both species, the contents (both as absolute levels and as percent compositions) of linoleic acid decreased and alpha-linolenic acid increased in tubers over the whole storage period, and possible mechanisms are discussed. Also, the absolute levels of these two acids were greater in S. phureja than in S. tuberosum, and this is discussed in relation to the development of flavor-related compounds during cooking. The polar lipids of one representative of S. tuberosum and of S. phureja were qualitatively similar. There were only minor differences in the polar lipid percentage compositions and in the corresponding fatty acid compositions of the individual polar lipids between the two species, although the absolute levels of the total, and of some individual, polar lipids were higher in S. phureja.     

Nitrogen and Potassium Fertilization Responses of Potato (Solanum tuberosum) cv. Spunta

A potato field experiment was conducted for 2 consecutive years to determine the effects of nitrogen (N) and potassium (K) fertilization rates on the yield and quality of potato cv. Spunta cultivated on soil low in N and K. A 3?×?4 complete factorial experiment was used with three rates of nitrogen (330, 495, and 660 kg N ha^–1) and four rates of potassium (112, 225, 450, and 675 kg K₂Ο ha^–1). An additional treatment without fertilization was used as the control. On soils low in N and K, potatoes showed low yield response to K fertilizer. The greatest tuber yields for both years were achieved at 495 kg N ha^–1 and 112 kg K₂O ha^–1 (29.81 t ha^–1) and 225 kg ha^–1 (27.13 t ha^–1), respectively. Differences in mean fresh weight due to treatment application were not significant. Application of 495 kg N ha^–1 significantly reduced harvest index (the ratio of tuber dry weight to the total dry weight at harvest) compared to 330 kg N ha^–1, but at 660 kg N ha^–1 harvest index achieved the greatest significant value. Potassium fertilization had no significant influence on harvest index. Nitrogen rates positively influenced the number of tubers. The addition of 450 kg K₂O ha^–1 significantly enhanced the number of tubers compared to the lower K rates, and the number was significantly decreased by the application of 675 kg K₂O ha^–1. Tuber dry-matter concentration was significantly promoted by N fertilization in both cultivation years, but it was negatively affected by K fertilization in the first year of cultivation. There was no change in tuber N with N application, but N application strongly increased nitrate (NO₃) concentration, which fluctuated between 360 and 1382 mg kg^–1 wet mass. Tuber NO₃ was negatively correlated with tuber yield, indicating that high levels of NO₃ in tubers can adversely affect yield. Tuber response to K fertilization was not in accordance with the rate of applied nutrient.     

Glycoalkaloid development during greening of fresh market potatoes (Solanum tuberosum L.)

Chlorophyll and glycoalkaloid synthesis in potato (Solanum tuberosum L.) tubers occur in direct response to light. The two processes are concurrent, but independent. Color photographic indices to subjectively grade fresh market potatoes for the extent of greening were developed under lighting conditions consistent with those of retail markets. Total glycoalkaloid (TGA) and chlorophyll accumulation for four cultivars were determined over the respective greening scales, thus calibrating the scales for TGA content. On average, TGA concentrations in complete longitudinal sections of tubers (flesh samples) were highest in Dark Red Norland followed by Russet Norkotah, Yukon Gold, and White Rose. TGA concentrations of flesh samples of White Rose and Yukon Gold tubers were somewhat variable and did not increase in direct proportion to greening level and chlorophyll content, particularly at higher levels of greening. TGA concentrations in Dark Red Norland and Russet Norkotah tubers were highly correlated (P < or = 0.001) with greening level and chlorophyll concentrations. When averaged over greening levels, skin samples contained 3.4- to 6.8-fold higher concentrations of TGAs than flesh samples, depending on the cultivar. The TGA concentration in periderm samples ranged from 37 to 160 mg/100 g of dry wt. Regardless of greening level, concentrations of TGAs in the flesh samples (including attached periderm) remained within limits presumed safe for human consumption. Discrimination of greened tubers on the basis of perceived glycoalkaloid toxicity is likely unfounded for the cultivars and greening levels studied.     

Effect of selenate supplementation on glycoalkaloid content of potato (Solanum tuberosum L.)

Potatoes (Solanum tuberosum L.) supplemented with increasing amounts of sodium selenate were analyzed for glycoalkaloid (GA) content. GAs were extracted with 5% acetic acid from freeze-dried tubers of two potato cultivars, Satu and Sini, harvested 10 weeks after planting as immature. The GAs alpha-solanine and alpha-chaconine were quantified by reverse-phase high-performance liquid chromatography (RP-HPLC) with diode array detection. Two independent experiments were performed. In the first experiment, the total GA concentration +/- standard error of the tubers ranged between 105 +/- 9 and 124 +/- 10 mg kg(-1) fresh weight in Satu and between 194 +/- 26 and 228 +/- 10 mg kg(-1) fresh weight in Sini. The ratio of alpha-solanine to alpha-chaconine was 0.2 in Satu and 0.5-0.6 in Sini. In the second experiment, the total GA concentration +/- standard error was 75 +/- 4 to 96 +/- 11 mg kg(-1) fresh weight, and the ratio of alpha-solanine to alpha-chaconine was 0.3-0.4 in Satu. A high sodium selenate supplementation (0.9 mg of Se kg(-1) quartz sand) slightly decreased the GA content in Satu, but this decrease was not statistically significant. Furthermore, at this addition level the Se concentration increased to a very high level of 20 microg g(-1) dry weight, which cannot be recommended for human consumption. In both experiments, the Se concentration in tubers increased with increasing sodium selenate application levels. Our results show that acceptable application levels of selenate did not have an effect on the GA concentration in immature potato tubers.     

Isolation and partial characterization of thermostable isoperoxidases from potato (Solanum tuberosum L.) tuber sprouts

Peroxidases (POD; EC 1.11.1.7) can cross-link cell wall polymers and may have an impact on the final textural quality of potato tubers. Because heat treatments are important during processing, the thermal properties of isoPODs from soluble and ionically and covalently bound fractions were studied from both potato tubers and sprouts. For both tissues, the ionically bound fraction was the most thermostable; approximately 20% of POD activity remained after a heat treatment of 10 min at 90 degrees C (for sprouts). The temperature profile of the ionically bound sprout fraction appeared to be nonlinear and suggested the presence of a very thermostable POD, which still showed activity after a heat treatment at 100 degrees C. Visualization by using isoelectric focusing confirmed the occurrence of a thermostable isoPOD with an IEP of 9.5, which displayed regeneration of activity after heat inactivation. This cationic POD was further purified by chromatography techniques, and by SDS-PAGE its molecular mass was estimated at 38 kDa.     

Andean potato cultivars (Solanum tuberosum L.) as a source of antioxidant and mineral micronutrients

Potato tubers were evaluated as a source of antioxidants and minerals for the human diet. A genetically diverse sample of Solanum tuberosum L. cultivars native to the Andes of South America was obtained from a collection of nearly 1000 genotypes using microsatellite markers. This size-manageable collection of 74 landraces, representing at best the genetic diversity among potato germplasm, was analyzed for iron, zinc, calcium, total phenolic, total carotenoid, and total vitamin C contents. The hydrophilic antioxidant capacity of each genotype was also measured using the oxygen radical absorbance capacity (ORAC) assay. The iron content ranged from 29.87 to 157.96 microg g-1 of dry weight (DW), the zinc content from 12.6 to 28.83 microg g-1 of DW, and the calcium content from 271.09 to 1092.93 microg g-1 of DW. Total phenolic content varied between 1.12 and 12.37 mg of gallic acid equiv g-1 of DW, total carotenoid content between 2.83 and 36.21 microg g-1 of DW, and total vitamin C content between 217.70 and 689.47 microg g-1 of DW. The range of hydrophilic ORAC values was 28.25-250.67 micromol of Trolox equiv g-1 of DW. The hydrophilic antioxidant capacity and the total phenolic content were highly and positively correlated (r = 0.91). A strong relationship between iron and calcium contents was also found (r = 0.67). Principal component analysis on the studied nutritional contents of the core collection revealed that most potato genotypes were balanced in terms of antioxidant and mineral contents, but some of them could be distinguished by their high level in distinct micronutrients. Correlations between the micronutrient contents observed in the sample and the genetic distances assessed by microsatellites were weakly significant. However, this study demonstrated the wide variability of health-promoting micronutrient levels within the native potato germplasm as well as the significant contribution that distinct potato tubers may impart to the intake in dietary antioxidants, zinc, and iron.     

Umami compounds are a determinant of the flavor of potato (Solanum tuberosum L.)

Vegetable flavor is an important factor in consumer choice but a trait that is difficult to assess quantitatively. The purpose of this study was to assess the levels of the major umami compounds in boiled potato tubers, in cultivars previously assessed for sensory quality. The free levels of the major umami amino acids, glutamate and aspartate, and the 5'-nucleotides, GMP and AMP, were measured in potato samples during the cooking process. Tubers were sampled at several time points during the growing season. The levels of both glutamate and 5'-nucleotides were significantly higher in mature tubers of two Solanum phureja cultivars compared with two Solanum tuberosum cultivars. The equivalent umami concentration was calculated for five cultivars, and there were strong positive correlations with flavor attributes and acceptability scores from a trained evaluation panel, suggesting that umami is an important component of potato flavor.     

Kunitz-type serine protease inhibitor from potato (Solanum tuberosum L. cv. Jopung)

An antifungal protein, AFP-J, was purified from tubers of the potato (Solanum tuberosum cv. L Jopung) by various chromatographic columns. AFP-J strongly inhibited yeast fungal strains, including Candida albicans, Trichosporon beigelii, and Saccharomyces cerevisiae, whereas it exhibited no activity against crop fungal pathogens. Automated Edman degradation determined the partial N-terminal sequence of AFP-J to be NH2-Leu-Pro-Ser-Asp-Ala-Thr-Leu-Val-Leu-Asp-Gln-Thr-Gly-Lys-G lu-Leu-Asp-Ala-Arg-Leu-. The partially sequence had 83% homology with a serine protease inhibitor belonging to the Kunitz family, and the protein inhibited chymotrypsin, pepsin, and trypsin. Mass spectrometry showed that its molecular mass was 13 500.5 Da. This protease inhibitor suppressed over 50% the proteolytic activity at 400 microg/mL. These results suggest that AFP-J is an excellent candidate as a lead compound for the development of novel antiinfective agents.     

Wounding stress increases the phenolic content and antioxidant capacity of purple-flesh potatoes (Solanum tuberosum L.)

Several abiotic stresses, including ethylene, methyl jasmonate, temperature, light, and wounding, were tested for their ability to induce accumulation of phenolic compounds and antioxidant capacity in purple-flesh potatoes (cv. All Blue). Results indicated that temperature, ethylene, methyl jasmonate, and light treatments did not significantly affect the accumulation of phenolic compounds compared to control samples. Only tubers with low initial anthocyanin levels treated with methyl jasmonate showed approximately 60% anthocyanin accumulation. Wounding induced the accumulation of phenolics compounds and an increase of PAL-activity in sliced tissue compared to the control. Total phenolics increased approximately 60% with a parallel 85% increase in antioxidant capacity. These results show that selection of appropriate abiotic stresses can enhance the nutritional and functional value of potatoes.     

黑龙江省马铃薯气候生产力特征及区划

利用黑龙江省81个气象站1975-2004年30 a的逐日气象资料及相应插值的网格同期逐日气象资料,采用WOFOST作物生长模型,模拟并分析了马铃薯气候生产力的空间分布特征,同时分析了各地气候生产力影响因素及分布特征;利用气候生产力的距平百分率、变异系数及与气候生产力密切相关的有关生育期的4个气候因子(平均气温、气温日较差、日平均日照时数、降水量),采用动态聚类分析方法,将黑龙江省马铃薯可能种植区初步划分为9类气候栽培区,为充分利用当地气候资源发展马铃薯生产提供参考。     

SSR标记的彩色马铃薯遗传多样性分析及指纹图谱构建

彩色马铃薯(指块茎的皮或肉为红、蓝、紫、橙色等)近年来日益为育种工作者所关注,很多彩色马铃薯品种(系)从形态学上难以鉴定是否为同一基因型,给育种工作带来诸多不便。本研究利用SSR标记对50份彩色马铃薯(Solanum tuberosumL.)材料进行了遗传多样性分析及指纹图谱构建。研究筛选出56对马铃薯SSR引物,对50份材料的基因组DNA进行PCR扩增,共检测出236个等位位点,其中多态性位点230个,多态性比率达97.46%。分析显示,基因型间遗传相似性系数在0.50~1.00之间。UPGMA聚类分析表明,在相似系数0.63处可将全部材料分为3大类。利用5对核心引物构建了50份供试材料的指纹图谱,并证明其属于44个基因型的,为彩色马铃薯资源鉴定和利用提供了依据。